The purpose of this proposal is to create the methodology for surveying the levels of the major class of cooking-induced mutagens, the aminoimidazoazaarenes (AIAs), in a variety of meats. We will also determine in mice exposed to AIAs the levels of these compounds in the body fluids. The AIA class of chemical includes IQ, MeIQ, MeIQx, and DiMeIQx which are potent mutagens in the Ames/Salmonella assay. Only one of these, IQ, has been tested in long-term bioassay, and it is carcinogenic. Determining the human health risk posed by these chemicals requires precise quantification of their levels in the major cooked meat classes, as well as determining their uptake and clearance rates. The ability to quantify these mutagens, which are often present only in part per billion concentrations is severely limited, using present analytical techniques. Current methods are limited both by lack of sensitivity and low sample throughout caused by the many steps in sample preparation. To improve sensitivity and throughput, we propose to develop and validate greatly improved analytical procedures for detection of these compounds based on immunoassays. We will develop a set of monoclonal antibodies first to AIAs and in later stages of the project to AIA-DNA adducts. Immunoassays offer several advantages over currently available analytical techniques in that very high specificity and sensitivity are possible with minimal sample cleanup, facilitating rapid sample throughout, and eventual automation. The immunoassays developed under this grant will greatly facilitate the quantification of AIA mutagens in food and fluids, allowing the necessary, comprehensive survey of meat sources and cooking conditions that is not possible with current methodology.